Project Summary Electronic BioSciences (EBS) will investigate and develop methodologies to accurately sequence microsatellite regions within the human genome to enable cancer genotyping. Microsatellites are simple/short repeats (1-10 nucleotides in length) that occur in tandem 5-50 times, and are among the most variable types of DNA sequence in the genome. Mutations to these microsatellite regions include expansion or contraction of the repeat number, single nucleotide polymorphisms (SNPs), and/or insertions or deletions (indels), which have been documented with cancer predisposition, cancer onset, and/or prognosis for many types of cancer, including colon, breast, bladder, brain, head and neck, skin, thyroid, kidney, prostate, lung, ovarian, and liver. To date, however, there is no technology presently available that is ideally suited for microsatellite sequencing/characterization, which has significantly limited the understanding of microsatellite mutations and the development of associated diagnostic/prognostic assays. During this two-year program, EBS will focus on developing the methodology to sequence and genotype microsatellites from colon cancer cell lines via direct, electronic, nanopore-based, single-molecule sequencing. The investigations performed during this program will enable new approaches to probe the human genome with unprecedented detail in order to relate and statistically characterize microsatellite sequences that signify a specific cancer type, predisposition, onset, and/or prognosis, directly improving basic cancer research and enabling eventual improved clinical diagnostics and/or prognostics technologies.